High-performance thin-layer chromatographic analysis with automated multiple development (AMD-HPTLC) of some commercial preparations and of laboratory-prepared extracts of Echinacea species is reported. Using an alkaline-based gradient system, hydrophilic and lipophilic compounds were separated in one chromatographic run. The chromatographic retention data were combined with spectroscopic data, i.e., UV and fluorescence spectra for the native compounds and after post-chromatographic derivatisation. Several isobutyramides and caffeic acid derivatives were identified. AMD-HPTLC fingerprint patterns were obtained for characterising and evaluating complex plant extracts.